title: DNA extraction and a cost-effective detection method for Echinococcus granulosus protoscoleces.
abstract: Most methods of DNA purification from protoscoleces of Echinococcus granulosus involve the use of expensive kits and may also require a second step after extraction for an effective purification. The present work describes an optimized cost-effective method that is fast and simple. This method is based on a chemical lysis with proteinase K with a subsequent one-step PCR detection. In this study we used already available primers and newly designed primers to amplify two fragments of different size corresponding to the mitochondrial cytochrome C oxidase subunit 1 gene. By one-step PCR, both fragments were successfully amplified from even a single protoscolex. This result demonstrates that this method of extraction is efficient even with small amounts of sample and that PCR is highly sensitive. The major advantage of this lysis-PCR method is that it avoids a second step of purification resulting in a simpler and more economical method. Our research will serve as a base for future studies on E. granulosus genotyping, mainly with wild mammals with a low number of cysts.
authors.affiliation: CONICET-Lab. Paleoparasitología y Arqueología Contextual, Depto. de Biología, Facultad de Ciencias, Exactas y Naturales, Universidad Nacional de Mar del Plata., Dean Funes 3350, Mar del Plata 7600, Buenos Aires, Argentina. Electronic address: rpetrigh@gmail.com.
authors.forename: R S
authors.initials: RS
authors.lastname: Petrigh
authors.forename: M H
authors.initials: MH
authors.lastname: Fugassa
mesh_terms: D000818:Animals; D003362:Cost-Benefit Analysis; D018509:DNA, Helminth / Q000235:genetics; D004443:Echinococcosis / Q000175:diagnosis*; D048209:Echinococcus granulosus / Q000235:genetics*; D003576:Electron Transport Complex IV / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000191:economics / Q000592:standards; D012680:Sensitivity and Specificity
chemical_list: D018509:DNA, Helminth; D003576:Electron Transport Complex IV
keywords: Echinococcus genotyping; One-step PCR; Single protoscolex lysis